Dwyer Amy R, Mouchemore Kellie A, Steer James H, Sunderland Andrew J, Sampaio Natalia G, Greenland Eloise L, Joyce David A, Pixley Fiona J
School of Medicine and Pharmacology, The University of Western Australia, Crawley, Western Australia, Australia.
School of Medicine and Pharmacology, The University of Western Australia, Crawley, Western Australia, Australia
J Leukoc Biol. 2016 Jul;100(1):163-75. doi: 10.1189/jlb.2A0815-344RR. Epub 2016 Jan 8.
A major role of colony-stimulating factor-1 is to stimulate the differentiation of mononuclear phagocytic lineage cells into adherent, motile, mature macrophages. The colony-stimulating factor-1 receptor transduces colony-stimulating factor-1 signaling, and we have shown previously that phosphatidylinositol 3-kinase p110δ is a critical mediator of colony-stimulating factor-1-stimulated motility through the colony-stimulating factor-1 receptor pY721 motif. Src family kinases are also implicated in the regulation of macrophage motility and in colony-stimulating factor-1 receptor signaling, although functional redundancy of the multiple SFKs expressed in macrophages makes it challenging to delineate their specific functions. We report a comprehensive analysis of individual Src family kinase expression in macrophage cell lines and primary macrophages and demonstrate colony-stimulating factor-1-induced changes in Src family kinase subcellular localization, which provides clues to their distinct and redundant functions in macrophages. Moreover, expression of individual Src family kinases is both species specific and dependent on colony-stimulating factor-1-induced macrophage differentiation. Hck associated with the activated colony-stimulating factor-1 receptor, whereas Lyn associated with the receptor in a constitutive manner. Consistent with this, inhibitor studies revealed that Src family kinases were important for both colony-stimulating factor-1 receptor activation and colony-stimulating factor-1-induced macrophage spreading, motility, and invasion. Distinct colony-stimulating factor-1-induced changes in the subcellular localization of individual SFKs suggest specific roles for these Src family kinases in the macrophage response to colony-stimulating factor-1.
集落刺激因子-1的一个主要作用是刺激单核吞噬细胞系细胞分化为贴壁、有运动能力的成熟巨噬细胞。集落刺激因子-1受体转导集落刺激因子-1信号，并且我们之前已经表明磷脂酰肌醇3激酶p110δ是通过集落刺激因子-1受体pY721基序介导集落刺激因子-1刺激的运动性的关键介质。Src家族激酶也参与巨噬细胞运动性的调节以及集落刺激因子-1受体信号传导，尽管巨噬细胞中表达的多种Src家族激酶存在功能冗余，使得难以描绘它们的具体功能。我们报告了对巨噬细胞系和原代巨噬细胞中单个Src家族激酶表达的全面分析，并证明了集落刺激因子-1诱导的Src家族激酶亚细胞定位变化，这为它们在巨噬细胞中的独特和冗余功能提供了线索。此外，单个Src家族激酶的表达既具有物种特异性，又依赖于集落刺激因子-1诱导的巨噬细胞分化。Hck与活化的集落刺激因子-1受体相关，而Lyn以组成型方式与该受体相关。与此一致，抑制剂研究表明，Src家族激酶对于集落刺激因子-1受体激活以及集落刺激因子-1诱导的巨噬细胞铺展、运动性和侵袭都很重要。单个Src家族激酶在集落刺激因子-1诱导下亚细胞定位的明显变化表明这些Src家族激酶在巨噬细胞对集落刺激因子-1的反应中具有特定作用。
